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Fig. 7. Oct4-deficient ZHBTc4 mESC are only partially rescued by
cPouV expression. ZHBTc4 cells transfected with mOct4,
XlPou91, cPouV or pou2 expression vectors were treated with
doxycyclin after selection of stable clones. AP-positive clones were obtained
with mOct4 (A), XlPou91 (B) and cPouV
(C), but no clones were isolated with empty control vector (empty) or
pou2. A rescue index (RI, the ratio between the number of clones in
the presence versus the absence of doxycyclin) of 1 is given in the presence
of mOct4 (D). This RI is the result of two independent
experiments with a total number of clones of 114/0, 82/19, 123/95, 74/8 and
82/0, respectively, in the absence/presence of doxycylin for the empty vector,
the mOct4, XlPou91, cPouV and pou2 expression vectors.
(E) Expression of pluripotency-associated genes in ZHBTc4 complemented
clones was analysed by real-time RT-PCR. A value of 1 was given to the level
detected in the mOct4-complemented clones. Nanog, Utf1,
Zfp42 (Rex1) and Tert expression was lower in clones
complemented by cPouV than in those complemented with
XlPou91. (F) ZHBTc4 cells were co-transfected in the presence
of luciferase reporter gene driven either by the 
PE promoter or the
Oct4 consensus sequence (ATGCAAAT). A value of 1 was given to the empty
vector. Each result is the average of four wells per condition, and two
independent experiments provided similar results. cPouV expression
activated both promoters.
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